The overall goal for this SHIFT SBIR application is to develop new, easy-to-use conjugatable adjuvant lipid vesicle (CALV) kits which contain Toll-like Receptor (TLR) agonists 3 and 7/8. Utilizing our proprietary VesiVax(R) CALV system and the Bacillus anthracis antigens, Protective Antigen (PA) and poly-3-D-glutamic acid (PGA), as model antigens, we will evaluate the adjuvant effect stimulated by candidate TLR3 and TLR7/8 agonists. We have already shown that the TLR4 agonist, MPL has an immunostimulatory effect and hypothesize that other TLR agonists will influence the immune response profile elicited to a particular target antigen and can be correlated with protective immunity against various pathogens. The rationale for testing liposomes containing different TLR agonists is to dissect the adjuvant effect of the TLR pathways by activating one or more of the receptors, and produce the optimal immune response for a target antigen of interest. We will evaluate in vitro and in vivo the immune responses elicited by the different TLR3 and TLR7/8 agonist formulations to our model antigens by characterizing antibody and cytokine production. The overall goal is to create an optimized set of kits that can be easily used by vaccinologists and immunologists to generate an effective immune response against the target antigen.